pBacPAK-His1 vector (V009315)

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V009315	pBacPAK-His1	In stock, 1 week for quality controls

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pBacPAK-His1

Antibiotic Resistance:: Ampicillin

Length:: 5414 bp

Type:: Baculovirus transfer vector

Replication origin:: ori

Source/Author:: Kitts PA.

Promoter:: polyhedrin

pBacPAK-His1 vector Map

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Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pBacPAK-His1 vector Sequence

LOCUS       40924_5659        5414 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Baculovirus transfer vector pBacPAK-His1, complete sequence, 
            beta-lactamase (bla) gene, complete cds.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5414)
  AUTHORS   Kitts PA.
  TITLE     pBacPAK-His1 transfer vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5414)
  AUTHORS   Kitts PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-APR-1996) Paul A. Kitts, CLONTECH Laboratories, Inc., 
            1020 East Meadow Circle, Palo Alto, CA 94303-4230, USA
REFERENCE   3  (bases 1 to 5414)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5414)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-APR-1996) Paul A. Kitts, CLONTECH Laboratories, Inc., 1020 East 
            Meadow Circle, Palo Alto, CA 94303-4230, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     This vector can be obtained from CLONTECH Laboratories, Inc., 1020 
            East  Meadow Circle, Palo Alto, CA 94303-4230, USA. To place an 
            order call  (415) 424-8222 or (800) 662-2566, extension 1. 
            International customers,  please contact your local distributor. For
            technical information, call  (415) 424-8222 or (800) 662-2566, 
            extension 3.
            This sequence has been compiled from information in the sequence 
            databases,  published literature and other sources, together with 
            partial sequences  obtained by CLONTECH. If you suspect there is an 
            error in this sequence,  please contact CLONTECH's Technical Service
            Department at (415) 424-8222 or  (800) 662-2566, extension 3 or 
            E-mail TECH@CLONTECH.COM.
FEATURES             Location/Qualifiers
     source          1..5414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     223..1050
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     promoter        1054..1145
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             1155..1172
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_recomb     1273..2630
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     rep_origin      2777..3232
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3514..3618
                     /label=AmpR promoter
     CDS             3619..4476
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4650..5238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"